Validation of a confirmatory analytical method for the determination of aflatoxins B₁, B₂, G₁ and G₂ in foods and feed materials by HPLC with on-line photochemical derivatization and fluorescence detection

A sensitive and selective analytical method for the simultaneous separation and quantitative determination of aflatoxins B1, B2, G1 and G2 in foodstuffs and materials for feed has been validated. The method is based on high performance liquid chromatography with on-line post-column photochemical derivatization and fluorescence detection. The chromatographic separation of aflatoxins was accomplished using a C18 column eluted with an isocratic mobile phase consisting of water, methanol and acetonitrile. The sample preparation required a simple extraction of aflatoxins with MeOH/H2O (80:20, v/v) and a purification step by immunoaffinity column cleanup. The total analysis time, including sample preparation and chromatographic separation, did not exceed 40 min with a run time of 10 min. The on-line photochemical derivatization ensures better results in terms of simplicity, sensitivity and reproducibility with respect to chemical derivatization techniques, and provides an increase of the peak resolution and an extent of automation in comparison with the electrochemical ones. The procedure for the determination of aflatoxins in food samples and cereals for animal consumption was extensively validated following Regulation (EC) No. 882/2004. Detection limits in wheat bran samples of 0.08 μg kg1 for AFB1, 0.02 μg kg1 for AFB2, 0.16 μg kg1 for AFG1 and 0.04 μg kg1 for AFG2 were attained. The method allows high recovery with mean values ranging from 72 to 94% and it satisfies the necessary requirements for sensitivity, linearity, selectivity, precision and ruggedness, demonstrating the conformity of the method with provisions of Regulation (EC) No. 401/2006.     

Comparison of methods and optimisation of the analysis of fumonisins B₁ and B₂ in masa flour, an alkaline cooked corn product

A comparison study of different extraction and clean-up procedures for the liquid chromatographic analysis of fumonisins B(1) (FB(1)) and B(2) (FB(2)) in corn masa flour was performed. The procedures included extraction (heat or room temperature) with acidic conditions or EDTA-containing solvents, and clean-up by immunoaffinity or C18 solid-phase extraction columns. Thereafter an analytical method was optimised using extraction with an acidic mixture of methanol-acetonitrile-citrate/phosphate buffer, clean-up through the immunoaffinity column and determination of fumonisins by liquid chromatography with automated pre-column derivatisation with o-phthaldialdehyde reagent. Recovery experiments performed on yellow, white and blue masa flours at spiking levels of 400, 800 and 1200 μg kg(-1) FB(1) and of 100, 200 and 300 μg kg(-1) FB(2) gave overall mean recoveries of 99% (±6%) for FB(1) and 88% (±6%) for FB(2). Good recoveries (higher than 90% for both FB(1) and FB(2)) were also obtained with corn tortilla chips. The limits of quantification of the method (signal-to-noise ratio of 10) were 25 μg kg(-1) for FB(1) and 17 μg kg(-1) for FB(2). The method was tested on different commercial corn masa flours as well as on white and yellow corn tortilla chips, showing fumonisin contamination levels (FB(1) + FB(2)) up to 1800 μg kg(-1) (FB(1) + FB(2)) in masa flour and 960 μg kg(-1) in tortilla chips. Over 30% of masa flours originating from Mexico exceeded the European Union maximum permitted level.     

Sources, vertical fluxes, and equivalent toxicity of aromatic hydrocarbons in coastal sediments of the Río de la Plata Estuary, Argentina

Settling particles and bottom sediments collected at 1, 2.5, and 4 km off the metropolitan Buenos Aires coast in the Río de la Plata were analyzed to evaluate the sources and toxicity of resolved (PAHs) and unresolved (AROUCM) aromatic hydrocarbons. PAHs (0003-2.1 microg g(-1)) and AROUCM (0.01-78 microg g(-1)) presented the highest concentrations nearthe Buenos Aires port and sewer and decreasing values up- and downstream and along on- and offshore gradients. Sediment traps deployed in the Central area revealed large aromatic fluxes (1.3 +/- 1.5 and 31 +/- 47 mg m(-2) day(-1) for PAHs and AROUCM). The composition of sedimentary PAHs was dominated by uniformly distributed high molecular weight pyrogenic PAHs (53 +/- 11% fluoranthene, pyrene, and heavier PAHs), followed by diagenetically derived perylene more abundant in less polluted sites (29 +/- 15%) and lower molecular weight petrogenic PAHs (18 +/- 7.1% phenanthrene, anthracene, and methylated compounds), which covaried inversely with perylene. PAH diagnostic ratios indicated a stronger influence of petrogenic discharges close to the shore and the prevalence of combustion of fossil fuels and vehicle emissions over wood in offshore sediments. Sediment cores showed sustained hydrocarbon levels with decreasing proportion of petrogenic PAHs and relative enrichment of pyrogenic components and perylene down to 20-cm depth. PAH toxicity assessment by sediment quality guidelines (SQG) and dioxin-equivalent factors (PAH TEQ: 0.08-395 pg g(-1) dw) identified 1-2.5 km sediments close to the port and sewer as the most affected area. According to SQG, dibenz[a,h]anthracene and pyrene were the most critical PAHs, followed by benzo[a]pyrene, benz[a]anthracene, and chrysene. In contrast, PAH TEQs were dominated by indeno[1,2,3-cd]pyrene, benzo[k]fluoranthene, benzo[a]pyrene, perylene, and benz[a]anthracene which accounted for an average 86 +/- 5.7% of total TEQs.     

Steam explosion improves extractability,antioxidant activity and α-glucosidase inhibitory activity of the constituents of Java tea (Clerodendranthus spicatus)

In this work, the steam explosion (SE) technology was exploited to extract valuable phytochemicals from Java tea (Clerodendranthus spicatus). Modification effects of SE on apparent morphology and physicochemical properties of Java tea were investigated to confirm that SE destroyed the cell wall structure and enhanced pore space, which facilitated the dissolution of ingredients. After SE pretreatment, the contents of phenolics, triterpenes, and sterols of Java tea extracts were maximally increased by 40.68%, 25.69%, and 40.11% in the leaves group, and increased by 147.09%, 204.50%, and 115.24% in the stems group, respectively. The antioxidant and α-glucosidase inhibition capacity of extracts were also obviously enhanced. Multivariate analysis indicated that phenolics and triterpenes were the main active substances in the extracts, and the leaves treated at 1.0 MPa and stems treated at 2.4 MPa showed higher comprehensive score. This study provided a promising method for Java tea pretreatment for improving extractability and bioactivity.     

Surprising formation of p-cymene in the oxidation of α-pinene in air by the atmospheric oxidants OH, O3, and NO3

Anthropogenic sources release into the troposphere a wide range of volatile organic compounds (VOCs) including aromatic hydrocarbons, whose major sources are believed to be combustion and the evaporation of fossil fuels. An important question is whether there are other sources of aromatics in air. We report here the formation of p-cymene [1-methyl-4-(1-methylethyl) benzene, C6H4(CH3)(C3H7)] from the oxidation of α-pinene by OH, O3, and NO3 at 1 atm in air and 298 K at low (<5%) and high (70%) relative humidities (RH). Loss of α-pinene and the generation of p-cymene were measured using GC-MS. The fractional yields of p-cymene relative to the loss of α-pinene, Δ [p-cymeme]/Δ [α-pinene], were measured to range from (1.6±0.2)×10(-5) for the O3 reaction to (3.0±0.3)×10(-4) for the NO3 reaction in the absence of added water vapor. The yields for the OH and O3 reactions increased by a factor of 4-8 at 70% RH (uncertainties are ±2s). The highest yields at 70% RH for the OH and O3 reactions, ～15 times higher than for dry conditions, were observed if the walls of the Teflon reaction chamber had been previously exposed to H2SO4 formed from the OH oxidation of SO2. Possible mechanisms of the conversion of α-pinene to p-cymene and the potential importance in the atmosphere are discussed.     

Macromolecular–structural interactions and phytochemical constituents of therapeutic herbs in pasta altered the functional properties,cooking profile and in vitro starch and protein digestibility

The current investigation was focused on developing and assessing the influence of different herbs (Ocimum sanctum, Withania somnifera, Bacopa monnieri and Asparagus racemosus) on bio-functional and structural characteristics of semolina-based pasta. Pasta were enriched with 2.5, 5, 7.5 and 10% herbs and significant difference (P < 0.05) in functional properties of blends, cooking quality of pasta, bioactive profile and antioxidant potential, in vitro nutrient digestibility and colour profile were noted. Herbs-enriched pasta showed modified functional properties and enhanced bioactive constituents. Modification in structural and molecular structures of control and 5% herbs-incorporated pasta was shown by Scanning Electron Microscopy and Attenuated Total Reflectance-Fourier Transform Infrared. Further, principal component analysis and agglomerative hierarchical clustering were employed to authenticate the difference in the incorporation of herbs and observations. The results suggest that herbs could be served as potential food components in valorised products of cereals.     

The effect of new bovine viral diarrhea virus introduction on somatic cell count,calving interval,culling, and calf mortality of dairy herds in the Dutch bovine viral diarrhea virus–free program

Bovine viral diarrhea virus (BVDV) infection has a major effect on the health of cows and consequently on herd performance. Many countries have implemented control or eradication programs to mitigate BVDV infection and its negative effects. These negative effects of BVDV infection on dairy herds are well documented, but there is much less information about the effects of new introduction of BVDV on dairy herds already participating in a BVDV control program. The objective of our study was to investigate the effect of a new BVDV introduction in BVDV-free herds participating in the Dutch BVDV-free program on herd performance. Longitudinal herd-level surveillance data were combined with herd information data to create 4 unique data sets, including a monthly test-day somatic cell count (SCC) data set, annual calving interval (CIV) and culling risk (CR) data sets, and a quarterly calf mortality rate (CMR) data set. Each database contained 2 types of herds: herds that remained BVDV free during the whole study period (defined as free herds), and herds that lost their BVDV-free status during the study period (defined as breakdown herds). The date of losing the BVDV-free status was defined as breakdown date. To compare breakdown herds with free herds, a random breakdown date was artificially generated for free herds by simple random sampling from the distribution of the breakdown month of the breakdown herds. The SCC and CIV before and after a new introduction of BVDV were compared through linear mixed-effects models with a Gaussian distribution, and the CR and CMR were modeled using a negative binomial distribution in generalized linear mixed-effects models. The explanatory variables for all models included herd type, BVDV status, year, and a random herd effect. Herd size was included as an explanatory variable in the SCC, CIV, and CMR model. Season was included as an explanatory variable in the SCC and CMR model. Results showed that free herds have lower SCC, CR, CMR, and shorter CIV than the breakdown herds. Within the breakdown herds, the new BVDV introduction affected the SCC and CMR. In the year after BVDV introduction, the SCC was higher than that in the year before BVDV introduction, with a factor of 1.011 [2.5th to 97.5th percentile (95% PCTL): 1.002, 1.020]. Compared with the year before BVDV breakdown, the CMR in the year of breakdown and the year after breakdown was higher, with factors of 1.170 (95% PCTL: 1.120; 1.218) and 1.096 (95% PCTL: 1.048; 1.153), respectively. This study reveals that a new introduction of BVDV had a negative but on average relatively small effect on herd performance in herds participating in a BVDV control program.     

Preparation,characterization and evaluation of the catalytic power of an hybrid compound [H3PMo12O40-chitosan] for the azoic-colored solutions

Inorganic–organic hybrid structures have become novel alternatives for many applications as they combine the advantages of both systems. In this current study, a hybrid compound [H₃PMo₁₂O₄₀-chitosan] was prepared using a wet impregnation method with different proportions (0-100 wt. %, considering the polymer as the main component). The chemical structure and the morphology of the synthesized compounds were studied using FT-IR and SEM. Their catalytic powers were evaluated for Naphtol Blue Black and Eriochrome Blue Black B. Experimental data revealed that the decolorization phenomenon depends on the structure of the dye it-self, the pH value, the initial dye concentration, the time contact, the oxidant dose and the temperature. Kinetic data, well described using the first order model, allowed us to establish the thermodynamic parameters. The calculated low values of Ea (7.48 and 8.14 KJ. mol-1) proved the efficiency of the prepared catalyst. Eriochrome Blue Black B could be decomposed more rapidly than Naphtol Blue Black. As example, after a period of 30 min of reaction, the target removals were, respectively, 70% and 15% for mono and bi-azoic dyes. The reuse of the catalyst is carried out and it was found to be recyclable more than three consecutives cycles.     

The contents of the neuro-excitatory amino acid β-ODAP (β-N-oxalyl-l-α,β-diaminopropionic acid), and other free and protein amino acids in the seeds of different genotypes of grass pea (Lathyrus sativus L.)

The free and protein amino acids of nine different genotypes of grass pea (Lathyrus sativus L.) seeds were analysed by HPLC with pre-column PITC (phenyl isothiocyanate) derivatisation. Among the free amino acids, homoarginine was quantitatively the most important (up to 0.8% seed weight) and stable while the neuro-excitatory amino acid β-ODAP (β-N-oxalyl-l-α,β-diaminopropionic acid) showed highest variation (0.02–0.54%) in the nine genotypes examined. Among protein amino acids, glutamic acid was quantitatively most significant, followed by aspartic acid, arginine, leucine, lysine and proline. The sulphur amino acid, methionine, showed the lowest concentration in all the L. sativus genotypes, and also in lentil (Lens culinaris) and in soybean (Glycine max) seeds analysed at the same time.